1. Expression of the mutants of HP1α and the mutant of SYCE2 in MCF7 cells or RPE cells.
Expression of the mutants of HP1α and the mutant of SYCE2 in MCF7 cells or RPE cells. (A) Exogenous expression of the HA-tagged wild-type HP1α protein and the HA-tagged mutants of HP1α in MCF7 cells. Western blot analysis shows successful knockdown of endogenous HP1α protein and expression of the exogenous HA-tagged wild-type HP1α protein and the HA-tagged mutants (V23M and KW41/42AA) of HP1α without affecting the expression levels of H3K9me3 in MCF7 cells. (B) Stable expression of the mutant SYCE protein lacking the hydrophobic sequence LTVLEGKS in RPE cells. For both A and B, 30 μg of total cell lysates was subjected to Western blotting analysis using the anti-HP1α antibody (A), anti-HA antibody (A), anti-H3K9me3 antibody (A), anti-FLAG antibody (B), and anti-CDK2 antibody (A, B). (C) Immunofluorescence visualization of the mutant FLAG-SYCE2 protein lacking the hydrophobic sequence LTVLEGKS stably expressed in RPE cells. Cells were stained with the anti-FLAG antibody (green) and DAPI (blue). Scale bar, 10 μm.
Noriko Hosoya et al. LSA 2018;1:e
© 2018 Hosoya et al.